BIOENERGETIC ASPECTS OF ALCOHOL CONSUMPTION

此申请是对ADAMHA研究科学家的申请 威廉·S·塞耶博士发展奖/二级 该奖项将允许塞耶博士将他的全部努力投入到研究中， 并将通过允许他 拓宽他在酒精相关研究方面的经验，学习 使用新的实验技术。后者将包括 免疫化学和核磁共振波谱。 这项拟议的研究将研究乙醇和 乙醇对线粒体膜组装和线粒体功能的影响 线粒体生物能量学。长期喂食酒精的大鼠将会 作为酒精性脂肪肝的动物模型。孤立 肝细胞，肝线粒体，磷酸化亚线粒体 颗粒和磷脂小泡用特定的 线粒体蛋白质，特别是细胞色素氧化酶，将是 用作实验系统。一种不活跃的细胞色素 氧化酶，最近通过免疫化学技术在 酒精中毒大鼠的亚线粒体颗粒将被分离出来 并以此为特征。血红素和蛋白质的生物周转 细胞色素氧化酶和其他线粒体的成分 体内的膜蛋白将从时间进程中确定 在特定的标记后放射性的降低 先驱物。乙醇对聚乙二醇单甲醚合成和组装的影响 细胞色素氧化酶和其他线粒体膜蛋白， 将通过免疫化学技术进行研究，使用分离的 肝细胞。线粒体膜蛋白的重组 进入磷脂囊泡将作为模型系统进行研究 膜组件。脂质成分与生理功能的关系 蛋白质含量和酶活性测定的性质 将探索重组囊泡的活性。的效果 酒精摄入对线粒体生物能量学和功能的影响 在肝细胞中将被研究通过检查 呼吸速率与电化学质子的关系 梯度和腺嘌呤核苷酸磷酸化系统。 乙醇对参与的质子转运反应的影响 电化学质子梯度的产生也将是 调查过了。这些研究将有助于理解 酒精消费和意志的生物能量后果 提供有关乙醇基本机制的见解 毒性。

This application is a request for an ADAMHA Research Scientist Development Award/Level II for William S. Thayer, Ph.D. The award will allow Dr. Thayer to devote his full efforts to research, and will enhance his scientific capabilities by allowing him to broaden his experience in alcohol-related research and learn the use of new experimental techniques. The latter will include immunochemistry and nuclear magnetic resonance spectroscopy. The proposed research will study the effects of ethanol and ethanol consumption on mitochondrial membrane assembly and mitochondrial bioenergetics. Rats fed ethanol chronically will be used as an animal model for alcoholic fatty liver. Isolated hepatocytes, liver mitochondria, phosphorylating submitochondrial particles, and phospholipid vesicles reconstituted with specific mitochondrial proteins, particularly cytochrome oxidase, will be used as experimental systems. An inactive form of cytochrome oxidase, recently identified by immunochemical techniques in submitochondrial particles from alcoholic rats, will be isolated and characterized. The biological turnover of heme and protein components of cytochrome oxidase, and other mitochondrial membrane proteins, in vivo will be determined from time courses of decreases in radioactivity following labeling with specific precursors. Effects of alcohol on the synthesis and assembly of cytochrome oxidase, and other mitochondrial membrane proteins, will be investigated by immunochemical techniques using isolated hepatocytes. Reconstitution of mitochondrial membrane proteins into phospholipid vesicles will be studied as a model system for membrane assembly. The role of lipid composition and physical properties in determining the protein content and enzymatic activity of reconstituted vesicles will be explored. Effects of ethanol consumption on mitochondrial bioenergetics and function in the liver cell will be studied by examining the regulatory relationships between respiration rate, electrochemical proton gradient and the adenine nucleotide phosphorylation system. Effects of ethanol on proton translocation reactions involved in generation of the electrochemical proton gradient will also be investigated. These studies will contribute to an understanding of the bioenergetic consequences of alcohol consumption and will provide insights concerning fundamental mechanisms of ethanol toxicity.